Digital microfluidic devices including dual substrates with thin-film transistors and capacitive sensing

ABSTRACT

An active matrix electrowetting on dielectric (AM-EWoD) device including a top substrate with driving electrodes and capacitive sensing. As depicted herein the bottom substrate includes a first plurality of electrodes to propel various droplets through a microfluidic region, while the top substrate includes a second plurality of electrodes that are configured to interrogate the droplets with capacitive sensing. In some embodiments, the top substrate has zones of high-resolution sensing and zones of low-resolution sensing.

RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.16/161,548, filed Oct. 16, 2018, published as U.S. Patent PublicationNo. 2019/0111433, which claims priority to U.S. Provisional PatentApplication No. 62/573,846, filed Oct. 18, 2017. All patents and patentapplications referenced in this specification are incorporated byreference in their entireties.

BACKGROUND

Digital microfluidic devices use independent electrodes to propel,split, and join droplets in a confined environment, thereby providing a“lab-on-a-chip.” Digital microfluidic devices are alternatively referredto as electrowetting on dielectric, or “EWoD,” to further differentiatethe method from competing microfluidic systems that rely onelectrophoretic flow and/or micropumps. A 2012 review of theelectrowetting technology was provided by Wheeler in “DigitalMicrofluidics,” Annu. Rev. Anal. Chem. 2012, 5:413-40, which isincorporated herein by reference in its entirety. The technique allowssample preparation, assays, and synthetic chemistry to be performed withtiny quantities of both samples and reagents. In recent years,controlled droplet manipulation in microfluidic cells usingelectrowetting has become commercially-viable; and there are nowproducts available from large life science companies, such as OxfordNanopore.

Most of the literature reports on EWoD involve so-called “passivematrix” devices (a.k.a. “segmented” devices), whereby ten to twentyelectrodes are directly driven with a controller. While segmenteddevices are easy to fabricate, the number of electrodes is limited byspace and driving constraints. Accordingly, it is not possible toperform massive parallel assays, reactions, etc. in passive matrixdevices. In comparison, “active matrix” devices (a.k.a. active matrixEWoD, a.k.a. AM-EWoD) devices can have many thousands, hundreds ofthousands or even millions of addressable electrodes. The electrodes aretypically switched by thin-film transistors (TFTs) and droplet motion isprogrammable so that AM-EWoD arrays can be used as general purposedevices that allow great freedom for controlling multiple droplets andexecuting simultaneous analytical processes.

Because of the restrictive requirements on the electric field leakage,most advanced AM-EWoD devices are constructed from polycrystallinesilicon (a.k.a. polysilicon, a.k.a. poly-Si). However, polysiliconfabrication is substantially more expensive than amorphous siliconfabrication, i.e., the type used in mass-produced active matrix TFTs forthe LCD display industry. Polysilicon fabrication processes are moreexpensive because there are unique handling and fabrication steps forworking with polysilicon. There are also fewer facilities worldwide thatare configured to fabricate devices from polysilicon. However, becauseof the improved functionality of polysilicon, Sharp Corporation has beenable to achieve AM-EWoD devices that include propulsion, sensing, andheating capabilities on a single active matrix. See, e.g., U.S. Pat.Nos. 8,419,273, 8,547,111, 8,654,571, 8,828,336, 9,458,543, all of whichare incorporated herein by reference in their entireties. An example ofa complex poly-Si AM-EWoD is shown in FIG. 1.

While poly-Si fabrication techniques allow implementation of complexAM-EWoD devices, the costs of poly-Si device production, combined with aglobal shortage of suitable fabrication facilities, has prevented theAM-EWoD technology from becoming widely available. There is a need fordifferent designs that can take advantage of existing amorphous siliconfabrication capacity. Such devices could be produced at lower cost, andin great quantities, making them ideal for commonplace diagnostictesting, such as immunoassays.

SUMMARY OF INVENTION

The invention addresses the shortcomings of the prior art by providingan alternate architecture for an AM-EWoD that is well-suited forconstruction from amorphous silicon substrates. In one instance, theinvention provides a digital microfluidic device, including a firstsubstrate, a second substrate, a spacer, and first and secondcontrollers. The first substrate includes a first plurality ofelectrodes coupled to a first set of thin-film-transistors, and includesa first dielectric layer covering both the first plurality of electrodesand the first set of thin-film-transistors. The second substrateincludes a second plurality of electrodes coupled to a second set ofthin-film-transistors, and includes a second dielectric layer coveringboth the second plurality of electrodes and the second set ofthin-film-transistors. The spacer separates the first and secondsubstrates, and creates a microfluidic region between the first andsecond substrates. The first controller is operatively coupled to thefirst set of thin-film-transistors and configured to provide apropulsion voltage to at least a portion of the first plurality ofelectrodes, while the second controller is operatively coupled to thesecond set of thin-film-transistors and configured to determine acapacitance between at least one of the second plurality of electrodesand a drive electrode. In some embodiments, the first dielectric layeris hydrophobic, and in other embodiments, the second dielectric layer ishydrophobic. In preferred embodiments, the first plurality of electrodesare arranged in an array, for example with at least 25 electrodes perlinear centimeter. In some embodiments, the second plurality ofelectrodes are interdigitated with the drive electrode. In someembodiments, a signal source is coupled to the drive electrode andconfigured to provide a time-varying voltage to the drive electrode. Insome embodiments the second substrate includes at least onelight-transmissive region, which may be, e.g., at least 10 mm² in area.The digital microfluidic device may be constructed from amorphous orpolysilicon.

In some embodiments, a digital microfluidic device will have two areasof different electrode densities, i.e., a high density (a.k.a.“high-res”) area, and a low density (a.k.a. “low-res”) area for thesensor electrode side. Such a design will allow a user to performparticle interrogation (i.e., capacitive sensing) to determinecomposition or size in one portion of the device, and then simplymonitor the location or presence of particles in another portion of thedevice. Overall, such a configuration simplifies the fabrication of adevice while also simplifying the data handling associated with thesensing functions.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 shows a prior art EWoD device including both propulsion andsensing on the same active matrix;

FIG. 2 depicts the movement of an aqueous-phase droplet between adjacentelectrodes by providing differing charge states on adjacent electrodes;

FIG. 3 shows a TFT architecture for a plurality of propulsion electrodesof an EWoD device of the invention;

FIG. 4 is a schematic diagram of a portion of the first substrate,including a propulsion electrode, a thin film transistor, a storagecapacitor, a dielectric layer, and a hydrophobic layer;

FIG. 5 is a schematic diagram of a portion of the second substrate,including a sensing electrode, a drive electrode, a thin filmtransistor, a dielectric layer, and a hydrophobic layer;

FIG. 6 shows a TFT architecture for a sensing electrode and a driveelectrode configured for capacitive sensing and evaluation ofmicrofluidic droplets;

FIG. 7 illustrates an embodiment wherein the sensing electrodes and thedrive electrode are interdigitated as part of the second substrate;

FIG. 8 illustrates a top view of a digital microfluidic device whereinthe sensing electrodes are arranged with varying regions of high and lowdensity. The electrode arrangement shown in FIG. 8 provides thenecessary functionality (droplet size determination and motion tracking)for many analytical functions while reducing the complexity of thedevice and the cost of production;

FIG. 9 illustrates an alternate embodiment including alight-transmissive region where droplets can be interrogated withelectromagnetic radiation, i.e., light. It is understood that both theprobe light and the resulting signal may enter/exit through the samelight-transmissive region;

FIG. 10 shows an alternative arrangement of sensing electrodes arrangedwith varying regions of high and low density;

FIG. 11 shows an alternative arrangement of sensing electrodes arrangedwith varying regions of high and low density;

FIG. 12 shows an alternative arrangement including elongated sensingelectrodes arranged with varying regions of high and low density;

FIG. 13 shows an alternative arrangement including elongated sensingelectrodes arranged with varying regions of high and low density.

DETAILED DESCRIPTION

As indicated above, the present invention provides an active matrixelectrowetting on dielectric (AM-EWoD) device including dual substrateswith thin-film transistors (TFT) and capacitive sensing. As depictedherein the “bottom” substrate includes a plurality of electrodes topropel various droplets through a microfluidic region. The “top”substrate includes a plurality of electrodes to provide a signal and todetect the presence and/or size and/or composition of a droplet withcapacitive sensing. The use of “top” and “bottom” is merely a conventionas the locations of the two substrates can be switched, and the devicescan be oriented in a variety of ways, for example, the top and bottomplates can be roughly parallel while the overall device is oriented sothat the substrates are normal to a work surface (as opposed to parallelto the work surface as shown in the figures). The top or the bottomsubstrate may include additional functionality, such as resistiveheating and/or temperature sensing. Because the devices incorporateTFT-based sensors, the devices have much higher sensitivity andresolution than known passive devices. Additionally, because both of theelectrodes needed for capacitive sensing are on the same substrate, thetop and bottom electrodes do not need to be aligned, and the sensingpixels can be of different sizes or configurations as compared to thepropulsion electrodes. Additionally, the designs can be implemented withamorphous silicon, thereby reducing the cost of production to the pointthat the devices can be disposable. It is also possible to use a-Si TFTsfor the bottom plate to benefit from their higher operation voltage, andpoly-Si TFTs on the top plate for higher sensitivity sensing.

The fundamental operation of an EWoD device is illustrated in thesectional image of FIG. 2. The EWoD 200 includes a cell filled with anoil 202 and at least one aqueous droplet 204. The cell gap is typicallyin the range 50 to 200 μm, but the gap can be larger. In a basicconfiguration, as shown in FIG. 2, a plurality of propulsion electrodes205 are disposed on one substrate and a singular top electrode 206 isdisposed on the opposing surface. The cell additionally includeshydrophobic coatings 207 on the surfaces contacting the oil layer, aswell as a dielectric layer 208 between the propulsion electrodes 205 andthe hydrophobic coating 207. (The upper substrate may also include adielectric layer, but it is not shown in FIG. 2). The hydrophobic layerprevents the droplet from wetting the surface. When no voltagedifferential is applied between adjacent electrodes, the droplet willmaintain a spheroidal shape to minimize contact with the hydrophobicsurfaces (oil and hydrophobic layer). Because the droplets do not wetthe surface, they are less likely to contaminate the surface or interactwith other droplets except when that behavior is desired.

While it is possible to have a single layer for both the dielectric andhydrophobic functions, such layers typically require thick inorganiclayers (to prevent pinholes) with resulting low dielectric constants,thereby requiring more than 100V for droplet movement. To achieve lowvoltage actuation, it is better to have a thin inorganic layer for highcapacitance and to be pinhole free, topped by a thin organic hydrophobiclayer. With this combination it is possible to have electrowettingoperation with voltages in the range +/−10 to +/−50V, which is in therange that can be supplied by conventional TFT arrays.

When a voltage differential is applied between adjacent electrodes, thevoltage on one electrode attracts opposite charges in the droplet at thedielectric-to-droplet interface, and the droplet moves toward thiselectrode, as illustrated in FIG. 2. The voltages needed for acceptabledroplet propulsion depend on the properties of the dielectric andhydrophobic layers. AC driving is used to reduce degradation of thedroplets, dielectrics, and electrodes by various electrochemistries.Operational frequencies for EWoD can be in the range 100 Hz to 1 MHz,but lower frequencies of 1 kHz or lower are preferred for use with TFTsthat have limited speed of operation.

As shown in FIG. 2, the top electrode 206 is a single conducting layernormally set to zero volts or a common voltage value (VCOM) to take intoaccount offset voltages on the propulsion electrodes 205 due tocapacitive kickback from the TFTs that are used to switch the voltage onthe electrodes (see FIG. 3). The top electrode can also have a squarewave applied to increase the voltage across the liquid. Such anarrangement allows lower propulsion voltages to be used for the TFTconnected propulsion electrodes 205 because the top plate voltage 206 isadditional to the voltage supplied by the TFT.

As shown in FIG. 3, an active matrix of propulsion electrodes can bearranged to be driven with data and gate (select) lines much like anactive matrix in a liquid crystal display. The gate (select) lines arescanned for line-at-a time addressing, while the data lines carry thevoltage to be transferred to propulsion electrodes for electrowettingoperation. If no movement is needed, or if a droplet is meant to moveaway from a propulsion electrode, then 0V will be applied to that(non-target) propulsion electrode. If a droplet is meant to move towarda propulsion electrode, an AC voltage will be applied to that (target)propulsion electrode.

The architecture of an amorphous silicon, TFT-switched, propulsionelectrode is shown in FIG. 4. The dielectric 408 must be thin enough andhave a dielectric constant compatible with low voltage AC driving, suchas available from conventional image controllers for LCD displays. Forexample, the dielectric layer may comprise a layer of approximately20-40 nm SiO₂ topped over-coated with 200-400 nm plasma-depositedsilicon nitride. Alternatively, the dielectric may compriseatomic-layer-deposited Al₂O₃ between 2 and 100 nm thick, preferablybetween 20 and 60 nm thick. The TFT is constructed by creatingalternating layers of differently-doped a-Si structures along withvarious electrode lines, with methods know to those of skill in the art.The hydrophobic layer 407 can be constructed from materials such asTeflon® AF (Sigma-Aldrich, Milwaukee, Wis.) and FlurorPel™ coatings fromCytonix (Beltsville, Md.), which can be spin coated over the dielectriclayer 408.

In the invention, a second substrate with TFT functionality isconstructed to provide capacitive sensing capabilities, and the twolayers are separated with a spacer that creates a microfluidic regionbetween the two layers. Capacitive sensing of droplets uses twoelectrodes, as shown in FIG. 6. Typically, an AC signal is applied to adriving electrode 506, whereby the AC signal produces acapacitively-coupled voltage on a nearby sensing electrode 505. Thecapacitively-coupled signal is measured by external circuitry, andchanges in the signal are indicative of the material between the driveelectrode 506 and the sensing electrode 505. For example, the coupledvoltage will be obviously different depending on whether oil 202 or anaqueous droplet 204 is between the electrodes because of the differencesin the relative permittivity between the materials. (Silicone oil has arelative permittivity of ε_(r)=2.5, ethanol has a relative permittivityof ε_(r)=24, and water has a relative permittivity of ε_(r)=80.)

The architecture of an amorphous silicon sensing layer, includingTFT-switched sensing electrodes 505 and drive electrodes 506 is shown inFIG. 5. The AC signal for the driving electrodes runs horizontally andonly one line at a time is activated to minimize capacitive coupling toread-out lines and “OFF” sensing electrodes. TFTs are not perfectswitches and have some small conductance even in the “OFF” state. Thismeans that a large number of OFF lines can have similar signal to one“ON” pixel. For this reason it is better to minimize capacitive signalsfrom ac voltages above and below the row being driven by only having acvoltages on the row being driven.

As shown in FIG. 6, the sensing and drive electrodes create a coplanargap cell. One major advantage is that the two plates do not need to beaccurately aligned, or even to have the same pixel pitch, thusfabrication of a two plate system is simplified. Additional details ofcapacitive sensing for droplets using interdigital gap cells can befound in, e.g., “Capacitance Variation Induced by Microfluidic Two-PhaseFlow across Insulated Interdigital Electrodes in Lab-On-Chip Devices”, TDong, C Barbosa, Sensors, 15, 2694-2708, (2015), which is incorporatedby reference in its entirety. The circuitry for detecting the capacitivesignals may include various electrical components, including amplifiers,multiplexing switches. Advanced designs may include an array of a-SiTFTs coupled to a multi-channel charge sensor, such as used for digitalx-ray imaging. See, “Front-end electronics for imaging detectors”, G. DeGeronimo, et al., Nuclear Instruments and Methods in Physics Research A,471 pp. 192-199, (2001), which is incorporated by reference in itsentirety.

In some embodiments, it is unnecessary to provide multiple independentdrive electrodes for the AC signal. As shown in FIG. 7, the driveelectrode can be arranged to be contiguous, but interdigitated with thesensing electrodes. (All of the electrodes shown in FIG. 7 are in thesame metal layer, but are shown in different colors to signify theirfunction.) In FIG. 7, the AC signal is provided to a singular drivingelectrode that runs horizontally across the surface, while varioussensing electrodes are “read” across the array. Typically, only onesensor line at a time is activated to minimize capacitive couplingbetween the AC signal from the driving electrode and sensing electrodesthat are in the “OFF” mode. Without such line-by-line readout, thesignal from the numerous sensing electrodes with a “null” state (e.g.,coupled to oil) will appear larger than proper, decreasing thesignal-to-noise of the correctly sensing electrodes. In an alternativeembodiment, the top substrate may include drive electrodes, sensingelectrodes, and an earthed grid. The drive and sensing electrodes can beused for droplet sensing, as described above, while the earthed gridprovides an electrode surface area opposite the propulsion electrodethat has low impedance to electrical ground.

The invention will use circuits coupled to the top drive and sensingelectrodes to provide capacitive sensing, thereby allowing the device totrack the position of droplets manipulated by the device. However, thesignal from capacitive sensing of droplets over a small sensingelectrode is also relatively small, thus one to three hundred lines ofsensor electrodes may be needed to obtain acceptable signal-to-noiseratios. Providing such a high density of sensing electrodes across theentire device would be expensive and unnecessary. Thus, for largerarrays (such as for combinatorial chemistry) it is preferred to havesmall localized areas with high densities of sensing pixels on the topplate for particle sizing, with lower density elsewhere for movementsensing.

As shown in FIG. 8, an AM-EWoD device can be created with differingdensities of sensing electrodes at various locations on the top plate.In the embodiment of FIG. 8, there are 200 dpi high-resolution areas onthe array for droplet size measurement, and 10 dpi resolution areas totrack droplet movement. In FIG. 8, the sensor would be 181.61 mm widefor 100 measurement lines. If the TFT EWoD propulsion substrate belowthe sensing plate had a uniform resolution of 200 dpi (electrodes perinch) then there would be 1430 rows of propulsion electrodes forcontrolling movement, mixing, etc. of droplets. In contrast, a devicelimited to one hundred sensing rows with a resolution of 180 dpi acrossthe entire device would only be 14.1 mm wide, resulting in only 111 rowsof propulsion electrodes; likely too small for complex assays. Thus, byproviding differing densities a larger device can be produced with allof the needed sensing capability. In general, a low-resolution area willinclude between 1 and 15 electrodes per linear centimeter, while ahigh-resolution area will include between 20 and 200 electrodes perlinear centimeter. Typically, the total area (length×width) of sensingelectrodes with the lower density (a.k.a., “low-res”) is greater thanthe total area of sensing electrodes with the higher density (a.k.a.,“high-res”). For example, there could be three times or greater of thelow-res area as compared to the high-res area s compared to the high-resarea. For example, there could be five times or greater of the low-resarea. For example, there could be ten times or greater of the low-resarea as compared to the high-res area.

An additional benefit of using different densities of sensing electrodesis that portions of the top plate can be provided with transparent, orotherwise light-transmissive, areas to allow further interrogation ofdroplets. For example, fluorescent markers may be observed byilluminating a droplet through the top substrate with a light source andthen using a detector and optionally color filters to observe theresulting fluorescence through the top substrate. In other embodiments,the light may pass through both the top and bottom substrates to allowabsorption measurements in the IR, UV, or visible wavelengths.Alternatively, attenuated (frustrated) total-internal reflectionspectroscopy can be used to probe the contents and or location ofdroplets in the system.

An embodiment of such a system is shown in FIG. 9, wherein a gap 910between sensing electrodes 905 is on the order of 2 mm, allowing light915 to pass from an objective 920 to illuminate a passing droplet 930.In an embodiment, the droplet 930 includes fluorescent molecules, andthe resulting fluorescent signal is collected back through the objective920 and split using a dichroic filter (not shown) to be detected with adetector (not shown). Thus, the design allows different types ofinformation, e.g., both capacitive and spectroscopic, to be collected ondroplets as they move through the system.

As discussed with respect to FIG. 8, the simplest way for implementinglow resolution sensing would be to have the same sensing pixel design asthe high-resolution areas, but have large spaces around the sensingpixel. This concept is illustrated in a different embodiment in FIG. 10.Using the design of FIG. 10, it would be possible for droplets to passbetween the low resolution sensing pixels, but a droplet controlalgorithm could be written to ensure droplets pass over the sensingpixels on a regular basis, allowing the size and composition of thedroplets to be monitored. As illustrated in FIG. 10, a uniformdistribution of low resolution pixels makes it possible to dramaticallyincrease the area over which sensing is available, while at the sametime allowing the use of commercially-available drivers. As analternative, the number of sensing pixels on any one vertical sensingline can be constant, while the sensing pixels are staggered, as shownin FIG. 11. Other patterns, such as pseudo-random may also be employedto maximize the interaction with the droplets, while reducing the actualnumber of sensing TFTs that must be fabricated and later addressed.

It is also possible to create low-resolution and high-resolution sensingareas using differently shaped electrodes, as shown in FIGS. 12 and 13.FIG. 12 shows square pixels in the high resolution sensing area andlarger rectangular sensing pixels in the low resolution sensing area.This design would be efficient for sensing movement up and down thearray, i.e., moving from one elongated electrode to another. This sametechnique could be implemented to make both horizontal and verticalelongated electrodes that would provide droplet tracking with lowerresolution. FIG. 13 shows low resolution area with vertical andhorizontal rectangular sensing pixels to detect vertical and horizontalmovement of droplets. Other geometric designs, such as spirals can alsobe used to facilitate location sensing with fewer electrodes and fewerTFTs. As shown in FIGS. 12 and 13, the droplets can be easily moved fromthe low density regions, where droplet creation, splitting, or mixingtake place, to high density regions where size and composition of thosedroplets can be evaluated.

From the foregoing, it will be seen that the present invention canprovide low-cost lab-on-a-chip functionality. In particular, by usingthe described architecture, an electrowetting on dielectric system canbe created using amorphous-silicon fabrication facilities and lower costdriving electronics. The invention makes efficient use of the availablesurfaces on both the top and the bottom of the EWoD device, but does notrequire alignment of the electrodes on the top and bottom surfaces.

It will be apparent to those skilled in the art that numerous changesand modifications can be made in the specific embodiments of theinvention described above without departing from the scope of theinvention. Accordingly, the whole of the foregoing description is to beinterpreted in an illustrative and not in a limitative sense.

1. A digital microfluidic device, comprising: a first substratecomprising a plurality of propulsion electrodes coupled to a set ofthin-film-transistors, and including a hydrophobic layer covering boththe plurality of propulsion electrodes and the set ofthin-film-transistors; a second substrate comprising a first pluralityof sensing electrodes and a first plurality of drive electrodes in afirst area of the second substrate, wherein the density of sensingelectrodes in the first area is between 20 and 200 sensing electrodesper linear centimeter; a second plurality of sensing electrodes and asecond plurality of drive electrodes in a second area of the secondsubstrate wherein the density of sensing electrodes in the second areais between 1 and 15 sensing electrodes per linear centimeter; a spacerseparating the first and second substrates and creating a microfluidicregion between the first and second substrates; a controller operativelycoupled to the set of thin-film-transistors and configured to provide apropulsion voltage between at least a portion of the propulsionelectrodes and the first plurality of drive electrodes and the secondplurality of drive electrodes.
 2. The digital microfluidic device ofclaim 1, wherein the set of thin-film-transistors comprises amorphoussilicon.
 3. The digital microfluidic device of claim 1, wherein theplurality of propulsion electrodes are arranged in an array.
 4. Thedigital microfluidic device of claim 3, wherein the array of propulsionelectrodes includes at least 25 propulsion electrodes per linearcentimeter.
 5. The digital microfluidic device of claim 1, wherein thesensing electrodes in the second plurality of sensing electrodes arebetween 0.01 and 5 mm in width.
 6. The digital microfluidic device ofclaim 1, wherein the second substrate includes at least onelight-transmissive region.
 7. The digital microfluidic device of claim6, wherein the light-transmissive region is at least 10 mm² in area. 8.The digital microfluidic device of claim 1, wherein the density ofsensing electrodes in the first area is at least three times as manysensing electrodes per 100 mm² as the density of sensing electrodes inthe second area.
 9. The digital microfluidic device of claim 1, whereinthe first area is smaller than the second area.
 10. The digitalmicrofluidic device of claim 9, wherein the second area is at leastthree times larger than the first area.
 11. The digital microfluidicdevice of claim 1, wherein the first plurality of sensing electrodes,the first plurality of drive electrodes, the second plurality of sensingelectrodes, and the second plurality of drive electrodes are all coveredby a second hydrophobic layer.
 12. The digital microfluidic device ofclaim 1, further comprising a dielectric layer between the hydrophobiclayer and the plurality of propulsion electrodes and the set ofthin-film-transistors.